TY - JOUR
T1 - Identification of immunogenic regions within the alternative reading frame protein of hepatitis C virus (genotype 3)
AU - Qureshi, H.
AU - Qazi, R.
AU - Hamid, S.
AU - Qureshi, S. A.
N1 - Funding Information:
Acknowledgments We are grateful to Dr. Najeeha Talat (Department of Pathology & Microbiology) and Mr. Junaid Iqbal and Ms. Hina Zuberi (Department of Biological & Biomedical Sciences) for their technical assistance. This work was funded by grants from the Higher Education Commission (HEC-20-342) and Aga Khan University Research Council (1UM/70139).
PY - 2011/9
Y1 - 2011/9
N2 - Hepatitis C virus (HCV) encodes ten classic proteins as well as a newly discovered alternative reading frame protein (ARFP) whose synthesis originates from the core region by a +1 frameshift. ARFP is produced by all HCV genotypes, but its function remains unknown. Although the immunogenicity of genotype 1- and 2- derived ARFP in infected hosts has been reported, no information is available for genotype 3-encoded ARFP. HCV genotype 3 core/ARFP region was PCR amplified, cloned, and sequenced. Recombinant ARFP and peptides were employed in ELISAs with patient serum samples. The effect of peptides on peripheral blood mononucleocytes (PBMCs) was also studied. DNA cloning and sequencing of HCV genotype 3 strain (PKHCV3) revealed it to encode 160 aa ARFP, which harbors a C-terminal extension of 36 aa. Serum from 74 of 88 patients (84%) contained rARFPreactive antibodies. Peptide ELISAs showed that all regions of rARFP were immunogenic, with peptide F7 (DSLSPRRAGAKAGPGLSPGT) being the most immunodominant. When incubated with PBMCs from HCVinfected individuals, F7 stimulated the production of TNFa and IL10. PKHCV3-derived ARFP encodes a 160 aa protein and antibodies against its entire length are found in 84% of all genotype 3-infected subjects. Peptide ELISAs revealed F7 to be highly immunogenic and capable of eliciting impressive T-cell responses.
AB - Hepatitis C virus (HCV) encodes ten classic proteins as well as a newly discovered alternative reading frame protein (ARFP) whose synthesis originates from the core region by a +1 frameshift. ARFP is produced by all HCV genotypes, but its function remains unknown. Although the immunogenicity of genotype 1- and 2- derived ARFP in infected hosts has been reported, no information is available for genotype 3-encoded ARFP. HCV genotype 3 core/ARFP region was PCR amplified, cloned, and sequenced. Recombinant ARFP and peptides were employed in ELISAs with patient serum samples. The effect of peptides on peripheral blood mononucleocytes (PBMCs) was also studied. DNA cloning and sequencing of HCV genotype 3 strain (PKHCV3) revealed it to encode 160 aa ARFP, which harbors a C-terminal extension of 36 aa. Serum from 74 of 88 patients (84%) contained rARFPreactive antibodies. Peptide ELISAs showed that all regions of rARFP were immunogenic, with peptide F7 (DSLSPRRAGAKAGPGLSPGT) being the most immunodominant. When incubated with PBMCs from HCVinfected individuals, F7 stimulated the production of TNFa and IL10. PKHCV3-derived ARFP encodes a 160 aa protein and antibodies against its entire length are found in 84% of all genotype 3-infected subjects. Peptide ELISAs revealed F7 to be highly immunogenic and capable of eliciting impressive T-cell responses.
UR - http://www.scopus.com/inward/record.url?scp=80054768583&partnerID=8YFLogxK
U2 - 10.1007/s10096-011-1194-1
DO - 10.1007/s10096-011-1194-1
M3 - Article
C2 - 21318731
AN - SCOPUS:80054768583
SN - 0934-9723
VL - 30
SP - 1075
EP - 1083
JO - European Journal of Clinical Microbiology and Infectious Diseases
JF - European Journal of Clinical Microbiology and Infectious Diseases
IS - 9
ER -
